Effect of different ozone treatments on aflatoxin degradation and physicochemical properties of pistachios

This study evaluated the efficiency of ozone for the degradation of aflatoxins in pistachio kernels and ground pistachios. Pistachios were contaminated with known concentrations of aflatoxin (AF) B₁, B₂, G₁ and G₂. Pistachio samples were exposed to gaseous ozone in a chamber at 5.0, 7.0 and 9.0 mg L^?1 ozone concentrations for 140 and 420 min at 20 °C and 70% RH. Aflatoxin degradation was determined by high performance liquid chromatography (HPLC). The efficiency of ozone for aflatoxin degradation in pistachios increased with increasing exposure time and ozone concentration. The results indicated that AFB₁ and total aflatoxins could be reduced by 23 and 24%, respectively, when pistachio kernels were ozonated at 9.0 mg L^?1 ozone concentration for 420 min. Only a 5% reduction in AFB₁ and total aflatoxin levels could be achieved for ground pistachios under the same conditions. No significant changes occurred in pH, color, moisture content and free fatty acid values of pistachio kernels and ground pistachios. Fatty acid compositions of pistachios did not change significantly after the ozonation treatments. No significant changes were found between sweetness, rancidity, flavor, appearance and overall palatability of ozonated and non‐ozonated pistachio kernels. Significant changes were observed in the organoleptic properties of ground pistachios, except rancidity, after 5.0 mg L^?1 ozone treatment for 140 min. Ozonation was found to be more effective for degrading aflatoxins in pistachio kernels than ground pistachios. Copyright © 2006 Society of Chemical Industry     

Effects of ozonation on microbiological,chemical and sensory attributes of vacuum-packaged rainbow trout stored at 4±0.5°C

The effects of ozone in aqueous solution on the shelf life of whole, vacuum-packaged rainbow trout, stored under refrigeration (4±0.5°C) were studied by monitoring the microbiological, chemical and sensory changes for a period of 15 days. Vacuum-packaged non-ozonated trout served as the control sample. Ozonation affected populations of bacteria namely, mesophilic aerobic bacteria, Pseudomonas spp. and H₂S-producing bacteria until day 11 of storage, Brochothrix thermosphacta, lactic acid bacteria and Enterobacteriaceae until day 8 of storage. Trimethylamine (TMA) values of all rainbow trout samples remained low (<3 mg N/100 g) until day 11 of storage, and then increased to 12.2, 8.9 and 4.7 mg N/100 g for the control and the samples ozonated for 60 and 90 min, respectively on day 15 of storage. Total volatile basic nitrogen (TVB-N) values remained relatively constant (20–25 mg N/100 g) until day 11 of storage, but increased to 61.1, 37.6 and 39.4 mg N/100 g flesh for the control and ozonated specimen for 60 and 90 min, respectively on day 15 of storage. Thiobarbituric acid (TBA) values remained relatively constant (1–3 mg MA/kg flesh) until day 12 of storage but increased to 8.4, 6.4 and 3.8 mg MA/kg flesh on day 15 of storage for the control and the ozonated trout for 60 and 90 min, respectively. Sensory evaluation (odor, taste and texture) of cooked rainbow trout showed a very good correlation with bacterial populations. On the basis of both sensory and microbiological data, a shelf life of 10 and 12 days was obtained for ozonated, vacuum-packaged and refrigerated rainbow trout at 60 and 90 min, respectively.     

Gamma irradiation as a means to eliminate Listeria monocytogenes from frozen chicken meat

Cells of Listeria monocytogenes ATCC 35152 were sensitive to gamma irradiation in phosphate buffer, pH 7.00 (D₁₀, dose required for 10% survival—0.15 kGy) at 0–5°C. The cells showed higher radiation survival when irradiated under frozen condition, with a D₁₀ of 0.3 kGy. The protection offered by shrimp/chicken/kheema homogenates (100 g litre^?1) was evidenced by even higher D₁₀ values (0.5 kGy) at both 0–5°C and cryogenic temperature. Boneless chicken meat samples were artificially inoculated with L monocytogenes ATCC 35152 cells at low (5 × 10³) colony-forming unit (cfu) g^?1 and high (5 × 10⁶ cfu g^?1) concentrations and irradiated at 1, 3, 4, 6 kGy doses under cryogenic conditions. The efficacy of the radiation process was evaluated by detecting L monocytogenes during storage at 2–4°C in the irradiated samples. These studies, when repeated with three other serotypes of L monocytogenes, clearly suggested the need for a dose of 3 kGy for elimination of 10³ cfu cells of L monocytogenes g^?1 from air-packed frozen chicken meat.     

INACTIVATION OF ESCHERICHIA COLI O157:H7 ON INOCULATED ALFALFA SEEDS WITH OZONATED WATER UNDER PRESSURE1

Alfalfa seeds inoculated with Escherichia coli O157:H7 (～10⁵ CFU/g) were subjected to low hydrostatic pressure. Seeds immersed in ozonated water at 4C were held at 8 and 12‐psi ozone pressure for 2, 4, 8, 16, 32, and 64 min. Alternatively, seeds were continuously sparged with ozone for up to 64 min and then held at 12 psi for 5 min. Controls consisted of sparging and pressurization with air. Thirty‐two minute treatments of continuous ozone sparging followed by pressurization of seeds at 12 psi for 5 min were repeated with the addition of four surfactants (Tween 20, Tween 80, SPAN 20, and SPAN 80) in the treatment water. Enumeration of E. coli O157:H7 on treated, untreated, and control seeds was done on tryptic soy agar supplemented with 50 μg/mL of nalidixic acid. The reduction in population of E. coli O157:H7 on seeds treated with the 8 and 12 psi hydrostatic pressure in ozonated water ranged from 0.74 ?1.56 log₁₀ CFU/g and 0.72 – 1.62 log₁₀ CFU/g, respectively. Control treatments carried out with air pressurization of seeds resulted in maximum population reductions of 1.55 log₁₀ and 1.83 log₁₀ CFU/g for 8 and 12 psi, respectively. For seeds treated with continuous ozone sparging (2 – 64 min) followed by pressurization at 12 psi for 5 min, the highest reduction was 2.03 log₁₀ CFU/g. Reductions were, however, not significantly different (P > 0.05) from control treatment (with air) which reduced the populations by 0.57 – 2.19 log₁₀ CFU/g. The presence of surfactants during continuous sparging of water followed by pressurization at 12 psi was not beneficial. None of the treatments adversely affected the germination of the seeds.     

Effect of ozonated water treatment on microbial control and on browning of iceberg lettuce (Lactuca sativa L.)

We examined the effect of ozonated water treatment on microbial control and quality of cut iceberg lettuce (Lactuca sativa L.). Fresh-cut lettuce was washed in ozonated water (3, 5, and 10 ppm) for 5 min at ambient temperature. The native bacterial population on the lettuce declined in response to a rise in ozone concentration. However, there was no further bacterial reduction (1.4 log CFU/g) above 5 ppm ozone. Although ozonated water treatment increased the phenylalanine ammonia lyase (PAL) activity of the lettuce stored at 10 degrees C compared with the water wash treatment after 1 day of storage, the concentration of ozone did not affect PAL activity. The a* value of the residue of the lettuce methanol extracts, which reflects the extent of browning, increased dramatically in lettuce treated with 10 ppm ozonated water compared with other treatments. Treatment with 3 or 5 ppm ozonated water resulted in more rapid changes in the a* value than after the water treatment. The combined treatment of hot water (50 degrees C, 2.5 min) followed by ozonated water (5 ppm, 2.5 min) had the same bactericidal effect as treatment with ozonated water (5 ppm, 5 min) or sodium hypochlorite (NaOCl, 200 ppm, 5 min), giving a reduction in bacteria numbers of 1.2 to 1.4 log CFU/g. The ascorbic acid content of the lettuce was not affected by these treatments. The combined treatment of hot water followed by ozonated water greatly inhibited PAL activity for up to 3 days of storage at 10 degrees C. Treatment with this combination greatly suppressed increases in the a* value, thus retarding the progress of browning compared with other treatments throughout the 6-day storage. NaOCl treatment also inhibited browning for up to 3 days of storage. Bacterial populations on the lettuce treated with sanitizers were initially reduced but then showed rapid growth compared with that of the water wash treatment, which did not reduce bacterial counts initially.     

Efficacy of Ozone Against Escherichia coli O157:H7 on Apples

Apples were inoculated with Escherichia coli O157:H7 and treated with ozone. Sanitization treatments were more effective when ozone was bubbled during apple washing than by dipping apples in pre‐ozonated water. The corresponding decreases in counts of E. coli O157:H7 during 3‐min treatments were 3.7 and 2.6 log₁₀ CFU on apple surface, respectively, compared to < 1 log₁₀ CFU decrease in the stem‐calyx region in both delivery methods. Optimum conditions for decontamination of whole apples with ozone included a pretreatment with a wetting agent, followed by bubbling ozone for 3 min in the wash water, which decreased the count of E. coli O157:H7 by 3.3 log₁₀CFU/g.     

Effect of dietary supplementation with vitamin E on characteristics of vacuum‐packed lamb

The effect of dietary vitamin E supplementation on lamb during vacuum‐packed storage was studied. Thirty‐six weaned male Manchego breed lambs were offered four dietary treatments (20, 270, 520 and 1020 mg vitamin E kg^?1 feed). Lambs were fed the vitamin E‐supplemented diet from 13 until 26 kg live weight. Pieces of M. longissimus dorsi were stored under vacuum at 2 ± 1 °C in the dark and meat quality was assessed after 5, 14 and 28 days of storage. Dietary supplementation significantly increased the α‐tocopherol concentration in the muscle (P < 0.001). Initially, lipid oxidation, meat colour and bacterial load were similar in all groups. In meat of non‐supplemented lambs the thiobarbituric acid reactive substance value increased throughout storage, whereas in meat of supplemented lambs it did not increase. Meat pigments and discolouration proportion were significantly affected by storage time (P < 0.001). The bacterial load was low initially, but after 28 days of storage it was close to 7 log₁₀ colony‐forming units (cfu) cm^?2 and Enterobacteriaceae surpassed the limit of acceptability of 2.5 log₁₀ cfu cm^?2, making the lamb unsuitable for human consumption. Meat of supplemented lambs displayed less lipid oxidation than that of their non‐supplemented counterparts, while meat colour and bacterial load were not affected by supplementation. Copyright © 2007 Society of Chemical Industry     

Ozone-high hydrostatic pressure synergy for the stabilization of refrigerated pitaya (Stenocereus pruinosus) juice

The synergistic effect of ozone (24 mg/O₃/L/min) and high hydrostatic pressure (HHP, 179–321 MPa) combination was evaluated with regard to individual effects of these over the microbial population in pitaya juice by using a response surface methodology and a repeated measures design with Tukey test. Treatment with 300 MPa/5 min reduced 6.89-log₁₀ and 0.89-log₁₀ CFU/mL of Saccharomyces cerevisiae and Listeria innocua population, respectively; and treatment with 9.6 min of ozonation time reduced 0.47-log₁₀ and 2.51-log₁₀ CFU/mL of S. cerevisiae and L. innocua population respectively; however, L. innocua was reduced 5.1-log₁₀ CFU/mL with exposure to ozone for 7 min followed by 316 MPa/5 min. Likewise, these treatment conditions kept native microbiota of the juice at non-detectable levels and achieved the highest sensory preference (79%) as compared to untreated juice at 30 d (5 ± 2 °C). Ozone-HHP reduced bacterial counts suggesting a microbiologically safe pitaya juice, contrary to what was obtained with individual application of these technologies.     

Efficacy of ozone in killing Listeria monocytogenes on alfalfa seeds and sprouts and effects on sensory quality of sprouts

A study was done to determine the efficacy of aqueous ozone treatment in killing Listeria monocytogenes on inoculated alfalfa seeds and sprouts. Reductions in populations of naturally occurring aerobic microorganisms on sprouts and changes in the sensory quality of sprouts were also determined. The treatment (10 or 20 min) of seeds in water (4 degrees C) containing an initial concentration of 21.8 +/- 0.1 microg/ml of ozone failed to cause a significant (P < or = 0.05) reduction in populations of L. monocytogenes. The continuous sparging of seeds with ozonated water (initial ozone concentration of 21.3 +/- 0.2 microg/ml) for 20 min significantly reduced the population by 1.48 log10 CFU/g. The treatment (2 min) of inoculated alfalfa sprouts with water containing 5.0 +/- 0.5, 9.0 +/- 0.5, or 23.2 +/- 1.6 microg/ml of ozone resulted in significant (P < or = 0.05) reductions of 0.78, 0.81, and 0.91 log10 CFU/g, respectively, compared to populations detected on sprouts treated with water. Treatments (2 min) with up to 23.3 +/- 1.6 microg/ml of ozone did not significantly (P > 0.05) reduce populations of aerobic naturally occurring microorganisms. The continuous sparging of sprouts with ozonated water for 5 to 20 min caused significant reductions in L. monocytogenes and natural microbiota compared to soaking in water (control) but did not enhance the lethality compared to the sprouts not treated with continuous sparging. The treatment of sprouts with ozonated water (20.0 microg/ml) for 5 or 10 min caused a significant deterioration in the sensory quality during subsequent storage at 4 degrees C for 7 to 11 days. Scanning electron microscopy of uninoculated alfalfa seeds and sprouts showed physical damage, fungal and bacterial growth, and biofilm formation that provide evidence of factors contributing to the difficulty of killing microorganisms by treatment with ozone and other sanitizers.     

Antimicrobial effects of pressurised carbon dioxide on Brochothrix thermosphacta in broth and foods

Antimicrobial effects of high‐pressure CO₂ on Brochothrix thermosphacta were investigated in a batch system. Inactivation rates increased with increasing pressure, temperature and exposure time. B thermosphacta suspended in physiological saline was completely inactivated under 6.05, 3.02 and 1.51 MPaCO₂ treatment at 35 °C after 10, 25 and 50 min respectively. Two phases were observed in the survival curves. The earlier stage was characterised by a slow rate of decrease in the number of B thermosphacta, which increased sharply at the later stage. B thermosphacta suspended in brain heart infusion broth was completely inactivated under 6.05 MPaCO₂ treatment after 80, 50 and 30 min at 25, 35 and 45 °C respectively. About 6.90 and 5.93 log cycles of B thermosphacta were reduced under 6.05 MPaCO₂ pressure at 45 °C in whole and skimmed milk respectively. The sterilisation effects of 6.05 MPaCO₂ pressure at 45 °C on both B thermosphacta and aerobic plate count were observed after 150 and 120 min respectively in skinned meat. © 2000 Society of Chemical Industry     

Quantitative assessment of the shelf life of ozonated apple juice

Sterile apple juice inoculated with S. cerevisiae ATCC 9763 (10³ CFU/mL) was processed in a bubble column with gaseous ozone of flow rate of 0.12 L/min and concentration of 33–40 μg/mL for 8 min. The growth kinetics of S. cerevisiae as an indicator of juice spoilage was monitored at 4, 8, 12 and 16 °C for up to 30 days. The kinetics was quantitatively described by the primary model of Baranyi and Roberts, and the maximum specific growth rate was further modeled as a function of temperature by the Ratkowsky type model. The developed model was successfully validated for the microbial growth of control and ozonated samples during dynamic storage temperature of periodic changes from 4 to 16 °C. Two more characteristic parameters were also evaluated, the time of spoilage of the product under static temperature conditions and the temperature quotient, Q ₁₀. At lower static storage temperature (4 °C), no spoilage occurred either for unprocessed or ozone-processed apple juice. In the case of ozone-processed apple juice, the shelf life was increased when compared with the controls, and the Q ₁₀ was found to be 7.17, which appear much higher than that of the controls, indicating the effectiveness of ozonation for the extension of shelf life of apple juice.     

Use of ozone in production chain of high moisture Mozzarella cheese

This work aimed to evaluate ozone effectiveness in reducing viable spoilage bacteria load throughout high moisture (HM) Mozzarella cheese-making process. At first, Mozzarella cheese samples were packaged with ozonated water (2 mg/L), stored at low temperature and monitored during shelf life. In a following phase cheese samples were put, before packaging, in direct contact with ozonated water at ozone concentrations of 2, 5 and 10 mg/L for 60 min. Then, gaseous ozone at concentrations of 10, 20 and 30 mg/m³ for different times was tested. In these experiments ozone was not effective in surface microbiological decontamination of cheeses. In all cases, there was no increase in the formation of primary and secondary lipid oxidation products.     

Growth or Survival of Listeria monocytogenes in Ready-to-Eat Meat Products and Combination Deli Salads During Refrigerated Storage

Growth or survival of Listeria monocytogenes in cold‐smoked salmon; sliced, cooked ham; sliced, roasted turkey; shrimp salad; and coleslaw obtained at retail supermarkets stored at 5 °C, 7 °C, or 10 °C (41 °F, 45 °F, or 50 °F, respectively) for up to 14 d was evaluated. Cold‐smoked salmon, ham, and turkey were obtained in case‐ready, vacuum packages. All food products were stored aerobically to reflect additional handling within the retail supermarket. Cold‐smoked salmon, ham, and turkey supported the growth of L. monocytogenes at all 3 storage temperatures. Fitted growth curves of initial populations (about 3 log₁₀ colony‐forming units [CFU]/g) in cold‐smoked salmon, ham, and turkey stored at 5 °C achieved maximal growth rates of 0.29, 0.45, and 0.42 log₁₀ CFU/g growth per day, respectively. Storage at 10 °C increased the estimated maximal growth rate of the pathogen by 0.56 to 1.08 log₁₀ CFU/ g growth per day compared with storage at 5 °C. A decline in populations of L. monocytogenes was observed in shrimp salad and coleslaw, and the rate of decline was influenced by storage temperature. Retention of viability was higher in shrimp salad than in coleslaw, where populations fell 1.2, 1.8, and 2.5 log₁₀ CFU/g at 5 °C, 7 °C, and 10 °C, respectively, after 14 d of storage. Inability of shrimp salad and coleslaw to support the growth of L. monocytogenes may be attributed to the acidic pH (4.8 and 4.5, respectively) of the formulations used in this study. Results show that the behavior of L. monocytogenes in potentially hazardous ready‐to‐eat foods is dependent upon the composition of individual food products as well as storage temperature.     

Development and Evaluation of an Ozonated Water System for Antimicrobial Treatment of Durum Wheat

ABSTRACT:  Ozonated water is reported to be effective in reducing the microbial load in foods such as fruits, vegetables, and grains. Ozonated water may be an effective alternative to chlorinated water in treating durum wheat before milling. Therefore, durum wheat was washed with ozonated water and analyzed for yeast and mold count (YMC) and aerobic plate count (APC). A system for producing and monitoring ozonated water was developed. The effect of water quality (tap, distilled, and ultra-pure), temperature (7, 15, and 25 °C), and pH (2, 4, and 6.5) was evaluated on the following: steady-state dissolved ozone concentration, ozone decay constant, half-life, mass transfer coefficient, equilibrium ozone concentration, and solubility ratio. The study of these parameters was important to attain a stable, high dissolved ozone concentration at the outset of washing and to have information for system improvement and scale-up. A 1% acetic acid solution (pH 2) at 15 °C resulted in high dissolved ozone concentration (21.8 mg/L) and long half-life (9.2 min). Subsequently, wheat was washed with 5 wash water types: distilled water, ozonated water (16.5 mg/L), chlorinated water (700 mg/L), acetic acid solution (1%), and acetic acid + ozonated water (1%, 20.5 mg/L). The treated samples were analyzed for YMC and APC. The acetic acid + ozonated water treatment was the most effective, with a reduction of 4.1 and 3.2 log₁₀ colony forming units/g in YMC and APC, respectively. Though ozonated water was not very effective alone, it was useful in combination with acetic acid.     

Efficacy of Chlorine Dioxide, Ozone, and Thyme Essential Oil or a Sequential Washing in Killing Escherichia coli O157:H7 on Lettuce and Baby Carrots

Chlorine dioxide (ClO₂), ozone, and thyme essential oil has been found to be effective in reducing pathogens, including Escherichia coli O157:H7, on selected produce. The efficacy of these sanitizers was evaluated, alone or through their sequential washing to achieve a 3 or more log reduction of mixed strains of E. coli O157:H7 on shredded lettuce and baby carrots. Samples sprinkle inoculated with mixed strains of E. coli O157:H7 were air-dried for 1 h at 22±2°C in a biosafety cabinet, stored at 4°C for 24 h, and then treated with different concentrations of disinfectants and exposure time. Sterile deionized water washing resulted in approximately 1log reduction ofE. coli O157:H7 after 10 min washing of lettuce and baby carrots. Gaseous treatments resulted in higher log reductions in comparison to aqueous washing. However, decolorization of lettuce leaves was observed during long exposure time. A logarithmic reduction of 1.48-1.97log₁₀ cfu/g was obtained using aqueous ClO₂ (10.0 mg/L for 10 min) ozonated water (9.7 mg/L for 10 min) or thyme oil suspension (1.0 mL/L for 5 min) on lettuce and baby carrots. Of the three sequential washing treatments used in this study, thyme oil followed by aqueous ClO₂/ozonated water, or ozonated water/aqueous ClO₂ were significantly (P<0.05) more effective in reducing E. coli O157:H7 (3.75 and 3.99log, and 3.83 and 4.34 log reduction) on lettuce and baby carrots, respectively. The results obtained from this study indicate that sequential washing treatments could achieve 3-4log reduction of E. coli O157:H7 on shredded lettuce and baby carrots.     

Inactivation of Lactobacillus fructivorans in physiological saline and unpasteurised sake using CO2 microbubbles at ambient temperature and low pressure

The ability of carbon dioxide microbubbles (MB‐CO₂) to inactivate Lactobacillus fructivorans suspended in physiological saline and unpasteurised sake at ambient temperature and a pressure lower than 2.0 MPa was investigated. The number of L. fructivorans cells in physiological saline solution containing 15% ethanol showed a 6‐log reduction following MB‐CO₂ treatment at 40 °C and 2.0 MPa for 50 min. The effectiveness of the treatment increased concomitantly with temperature, pressure and ethanol concentration of the sample solution but was unaffected by the glucose concentration in the sample solution. Furthermore, the number of L. fructivorans cells showed a 5‐log reduction in sake after MB‐CO₂ treatment at 40°C and 2.0 MPa for 60 min. Sensory evaluation revealed no significant difference between MB‐CO₂‐treated sake and unpasteurised sake. These results indicated that MB‐CO₂ treatment was highly effective for the inactivation of L. fructivorans and might become a practical method for pasteurising sake at ambient temperature.     

Microbiological Studies of the Cause of Discoloration of Common Dolphin (Coryphaena hippurus) Fillets

The effect of killing method of common dolphins aboard vessels on green pigment formation, and the microbiological survey of the green meat were studied. Eight bacterial strains which produced H₂S in peptone iron agar at both 4 and 25°C were isolated from green meat samples. They constituted one strain of Alteromonas putrefaciens and seven strains of Proteus vulgaris. Discoloration occurred when each of the eight isolates was reinoculated in normal fillets at 4°C. The bleeding operation has little effect on reducing green discoloration of the dolphen meat. Discoloration of the meat could be prevented by icing the fish and keeping them cold until they could be filleted and frozen.     

Thermal Inactivation of the Kanagawa Hemolysin of Vibrio parahaemolyticus in Buffer and Shrimp

A semipurified thermostable direct hemolysin (Kanagawa hemolysin) was prepared from a clinical isolate of Vibrio parahaemolyticus (strain 533- 72). Preliminary experiments determined that the hemolytic activity and mouse lethality of the hemolysin were inactivated at the same rate, that the rate was linear, and that the pH range of optimum thermal stability was pH 5.5-6.5. When approximately 256 rat erythrocyte units/g of Kanagawa hemolysin were heated at 115°, 120°, 125°, and 130°C, D values ranged from 46.5-13 min (z_c= 27°C) in Tris buffer and 48.1-10.4 min (z_c= 23.2°C) in shrimp homogenate, both at pH 7.0.     

Effects of ozone and chlorine postharvest treatments on quality of fresh‐cut red bell peppers

The effects of chlorine (200 μL L^?1), ozonated water (1 μL L^?1) and gaseous ozone (0.7 μL L^?1) on physicochemical attributes and microbial quality of minimally processed red bell peppers were studied. In all the experiments, O₂ continuously decreased and CO₂ concentration increased, the pH augmented and a significant softening was observed in all the fruits. By day 14, L* values decreased in all the fruits, with the greatest changes found in the chlorinated samples (approximately 12 units). Peppers treated with the aqueous solutions showed greater changes in the quality attributes with increasing washing times and especially when chlorine was used. The exposure for three min to gaseous O₃ reduced the mesophiles, psychrotrophes and fungal populations of the fresh‐cut peppers in 2.5, 3.3 and 1.8 log units, respectively. Combined with modified atmosphere, this could be an appropriate method to maintain the quality and extend the storage period of minimally processed red bell peppers.     

Comparison of the Proteolytic Activities of New Commercially Available Bacterial and Fungal Proteases toward Meat Proteins

The hydrolytic activity of 3 commercially available protease preparations (bacterial protease G, fungal 31000, and fungal 60000) were examined using fluorescent‐labeled casein, azo dye‐impregnated collagen, and meat protein extracts from bovine M. semimembranosus and Achilles tendon, and compared to that of papain. Assays showed that all proteases exhibited little activity at low temperature (5 °C), and maximal activity at 45 °C. The pH, at which optimal activity was observed for each of the protease preparations, differed and ranged from pH 5.0 to 8.0. Kinetic parameters (K_M and V_max) were also different between protease preparations, with the bacterial protease G and papain exhibiting significantly higher V_max values (P < 0.001) and lower K_M values (P < 0.01) for the casein substrate than the 2 fungal protease preparations. Meat protein hydrolysis was displayed on SDS‐PAGE and proteins analyzed with mass spectrometry. The protease preparations were shown to have varying affinity toward different meat proteins. The bacterial protease G preparation was efficient at hydrolyzing most myofibril and collagen proteins, and appeared to be more efficient than papain at hydrolyzing collagen proteins. On the other hand the 2 fungal protease preparations showed a selective specificity toward meat myofibrillar proteins, and the fungal 60000 protease preparation exhibited high affinity toward collagen γ and collagen type I chain B proteins. The results generated in this study demonstrated that these commercial proteases have good potential for use in meat tenderization applications due to their mild and complementary effects on different meat proteins. Practical Application : Bacterial and fungal protease preparations exhibited varying affinities for hydrolyzing meat proteins. This selective moderate capability of microbial proteases compared to papain is potentially an advantage in avoiding over‐tenderization in meat. On the other hand, the bacterial protease G preparation, which appeared to be more efficient at hydrolyzing connective tissue proteins than papain, could be beneficial in tenderizing meat with high connective tissue content. The synergistic effect of these protease preparations could be incorporated into a meat tenderizing formula to give the tenderizer a broad activity spectrum, thus able to target different cuts of meat.